WO2012113835A1 - Composés d'ellagitanin c-glucosidique destinés à être utilisés pour modifier l'arrangement supramoléculaire de l'actine et pour le traitement de l'ostéoporose, du cancer, d'une infection bactérienne et une d'infection virale - Google Patents

Composés d'ellagitanin c-glucosidique destinés à être utilisés pour modifier l'arrangement supramoléculaire de l'actine et pour le traitement de l'ostéoporose, du cancer, d'une infection bactérienne et une d'infection virale Download PDF

Info

Publication number
WO2012113835A1
WO2012113835A1 PCT/EP2012/053017 EP2012053017W WO2012113835A1 WO 2012113835 A1 WO2012113835 A1 WO 2012113835A1 EP 2012053017 W EP2012053017 W EP 2012053017W WO 2012113835 A1 WO2012113835 A1 WO 2012113835A1
Authority
WO
WIPO (PCT)
Prior art keywords
actin
compound
vescalagin
glucosidic
roburin
Prior art date
Application number
PCT/EP2012/053017
Other languages
English (en)
Inventor
Stéphane QUIDEAU
Elisabeth GENOT
Frédéric SALTEL
Céline DOUAT-CASASSUS
Daniela Mélanie DELANNOY LOPEZ
Original Assignee
Institut National De La Sante Et De La Recherche Medicale (Inserm)
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Institut National De La Sante Et De La Recherche Medicale (Inserm) filed Critical Institut National De La Sante Et De La Recherche Medicale (Inserm)
Priority to US14/000,766 priority Critical patent/US20130324595A1/en
Priority to EP12704839.5A priority patent/EP2678012A1/fr
Publication of WO2012113835A1 publication Critical patent/WO2012113835A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/357—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having two or more oxygen atoms in the same ring, e.g. crown ethers, guanadrel
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365—Lactones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention concerns a C-glucosidic ellagitannin compound, or a 5 metabolite, for use for altering the supramolecular arrangement of actin in an individual in need thereof. It also pertains to pharmaceutical compositions comprising a C-glucosidic ellagitannin compound and/or metabolites thereof and one or more physiologically acceptable carriers. It finally concerns a C-glucosidic ellagitannin compound or a metabolite thereof, optionally detectably labeled, for in vitro use as a tool for studying 10 cellular mechanisms involving actin, or for detecting F-actin in a cell.
  • Cancer is an unregulated proliferation of cells due to loss of normal controls, resulting in unregulated growth, lack of differentiation, local tissue invasion, and often, metastasis. Cancer can develop in any tissue or organ at any age. Many cancers are curable if detected at an early stage, and long-term remission is often possible in later stages. However, cure is not always possible and is not attempted in some advanced 0 cases. The development of drugs effective against cancer and having limited toxic side effects thus remains a critical need.
  • tumor angiogenesis factors are produced to promote formation of the vascular supply required for further tumor growth.
  • a tumor may shed cells into the circulation. Although most circulating tumor cells die as a result of intravascular trauma, an occasional cell may adhere to the vascular endothelium and penetrate into surrounding tissues, generating independent tumors (metastases) at distant sites. Metastatic tumors grow in much the same manner as primary tumors and may subsequently give rise to other metastases.
  • adhesion to the extracellular matrix and to neighbouring cells 5 plays a central role in the control of cell survival, growth, differentiation, motility, and tissue integrity.
  • Oncogenic transformation profound changes occur in the organization of the actin cytoskeleton, manifesting on cell morphology and motility.
  • Increased proliferation a hallmark of cancer cells, is highly dependent upon actin dynamics and cell adhesion.
  • Adhesive interactions involve specialized transmembrane receptors that are linked to the cytoskeleton through junctional plaque proteins.
  • actin-binding proteins including a-actinin, vinculin, tropomyosin and profilin.
  • Osteoporosis is a progressive metabolic bone disease that decreases bone density with deterioration of bone structure. Skeletal weakness leads to fractures with minor or inapparent trauma, particularly in the thoracic and lumbar spine, wrist, and hip. Acute or chronic back pain is common. Prevention and treatment involve calcium and vitamin D supplements, exercises to maximize bone and muscle strength and minimize the risk of falls, and drug therapy to preserve bone mass or stimulate new bone formation.
  • Osteoblasts cells that make the organic matrix of bone and then mineralize bone
  • osteoclasts cells that resorb bone
  • PTH parathyroid hormone
  • calcitonin calcitonin
  • estrogen calcitonin
  • vitamin D calcitonin
  • cytokines cytokines
  • Bone mass in men and women occurs by the mid 20s. Bone mass plateaus for about 10 yr, during which time bone formation approximately equals bone resorption. After this, bone loss occurs at a rate of about 0.3 to 0.5% per year. Beginning with menopause, bone loss accelerates in women to about 3 to 5% per year for about 5 to 7 year.
  • the major mechanism is increased bone resorption, which results in decreased bone mass and microarchitectural deterioration, even though other mechanisms also contribute to osteoporosis.
  • the mechanisms of bone loss may involve local changes in the production of bone-resorbing cytokines (such as increases in cytokines that stimulate bone resorption), impaired formation response during bone remodeling (probably caused by age-related decline in the number and activity of osteoblasts), and other factors such as a decline in local and systemic growth factors.
  • the goals of treatment against osteoporosis are to preserve bone mass, prevent fractures, decrease pain, and maintain function.
  • the rate of bone loss can be slowed with drugs (e.g. bisphosphonates or other anti-resorptive drugs) and, when possible, modification of risk factors.
  • drugs e.g. bisphosphonates or other anti-resorptive drugs
  • Calcium and vitamin D intake and physical activity must be adequate for drug treatment to be effective.
  • Bisphosphonates are first-line drug therapy. By inhibiting bone resorption, bisphosphonates preserve bone mass and can decrease vertebral and hip fractures by 50%. All increase bone mineral density and decrease risk of at least vertebral fractures.
  • Risk factors also include bisphosphonate use and cancer.
  • Bisphosphonates may further be associated with atrial fibrillation, but the mechanism is not clear and there has been no association with increased cardiovascular mortality.
  • Estrogen can preserve bone density and prevent fractures. Most effective if started within 4 to 6 yr of menopause, estrogen may slow bone loss and possibly reduce fractures even when started much later. However, use of estrogen increases the risk of thromboembolism and endometrial cancer and may increase the risk of breast cancer.
  • the risk of endometrial cancer can be reduced in women with an intact uterus by taking a progestin with estrogen.
  • taking a combination of a progestin and estrogen increases the risk of breast cancer, coronary artery disease, stroke, and biliary disease.
  • PTH which stimulates new bone formation
  • the osteoclast is the specialized cell that is responsible for bone resorption. It is a highly polarized cell that must adhere to the bone surface, where it undergoes alternative cycles of migration and resorption. Actin reorganization is critical for both processes. Osteoclast motility is mediated by podosomes, which are highly dynamic F-actin structures. Resorbing osteoclasts form a related actin complex, the sealing zone, which provides the boundary for the resorptive microenvironment. Similar to podosomes, the sealing zone is highly dependent on actin dynamics to allow efficient resorption. The integrity of the supramolecular arrangement of actin also plays a major role in the formation of the osteoclastic actin ring, a prerequisite for bone resorption.
  • the ellagitannins are a class of hydrolysable tannins formed when gallic acid, a phenol monomer, esterifies with the hydroxyl groups of a polyol carbohydrate such as glucose and oxidatively couples into hexahydroxydiphenoyl (HHDP) units, from which ellagic acid can be hydrolytically released.
  • HHDP hexahydroxydiphenoyl
  • C-qlucosidic ellagitannins refers to a particular group of ellagitannins, essentially occurring in plant species of only three subclasses of the Cronquist angiosperm classification (i.e., Hamamelidae, Rosidae and Dilleniidae), that comprises a very unique series of highly hydrosoluble C-glucosidic variants in that the usual glucopyranose core is replaced by a rarely encountered-in-nature open-chain glucose resulting from the establishment of their C-aryl glucosidic bond.
  • Several of these C-glucosidic ellagitannins further display a nonahydroxyterphenoyl (NHTP) unit triply connected at positions 2, 3 and 5 of their glucose core ( Figure 1 ).
  • NHTP nonahydroxyterphenoyl
  • the inventors have described for the first time the effects of several C-glucosidic ellagitannin on cellular actin, one of the most abundant structural proteins in eukaryotic cells (Example 1 ).
  • Monomeric globular actin (G-actin) subunits assemble via an ATP- dependent process into polymeric fibrillar actin (F-actin) filaments that are further ordered into three-dimensional architectures by interacting with so-called actin-binding proteins (ABPs) to establish the functional actin cytoskeleton.
  • ABSPs actin-binding proteins
  • the expression "supramolecular arrangement of actin” refers to such three-dimensional architectures of polymeric F-actin filaments.
  • a dynamic equilibrium between the G-actin and F-actin states continuously ensures the adaptation of the actin cytoskeleton during its various implications in determining and/or controlling inter alia cell shape, cytokinesis, motility, adhesion and gene expression.
  • the inventors have surprisingly found that the C-glucosidic ellagitannin vescalagin interacts with F-actin, and alters the supramolecular arrangement of actin by winding the actin filaments into fibrillar aggregates (Examples 5 and 6).
  • the inventors have also found that the C-glucosidic ellagitannin vescalagin is capable of crossing the plasma membrane, and that it specifically binds polymeric F-actin, thus interfering with its function, but not with micro-tubules. Furthermore, they have found that it does not induce depolymerization of F-actin, but rather promotes the polymerization of F-actin, thus displacing the G-actin/F-actin equilibrium in favour of F-actin.
  • a first aspect of the invention is a C-glucosidic ellagitannin compound and/or a metabolite thereof for use for altering the supramolecular arrangement of F-actin in an individual in need thereof.
  • the C-glucosidic ellagitannins consist of vescalagin, vescalin, castalagin, castalin, grandinin, roburin A, roburin B, roburin C, roburin D, and roburin E.
  • the compound for use for altering the supramolecular arrangement of actin in an individual in need thereof is selected from the group consisting of vescalagin (CAS 36001 -47-5), vescalin, castalagin (CAS 24312-00-3), and castalin (CAS 19086-75-0), and metabolites thereof.
  • the metabolite of the C-glucosidic ellagitannin according to the invention may refer to any intermediate or product of the C-glucosidic ellagitannin metabolism, such as:
  • - ellagic acid which may be derived from the hydrolysis of native C-glucosidic ellagitannins such as vescalagin, castalagin, grandinin and roburins A-E,
  • urolithin A urolithin B
  • urolithin C urolithin D
  • urolithin D which may notably be derived from gradual metabolism of ellagic acid by the intestinal microbiota, or
  • - conjugates thereof such as glucosides or glucuronides of urolithin A, of urolithin B, of urolithin C, of urolithin D, of ellagic acid or of dimethyl ellagic acid.
  • the C-glucosidic ellagitannins or the metabolites thereof according to the invention may be obtained by extraction from oak or chestnut wood.
  • vescalin and castalin may also be produced by hemi-synthesis from vescalagin and from castalagin, respectively, in particular with concomitant production of ellagic acid, as described by the inventors in Quideau et al., Chem. Eur. J. 2005, 1 1 , 6503 - 6513.
  • altering / alteration of the supramolecular arrangement of actin is meant any kind of modification of the supramolecular arrangement of actin.
  • the supramolecular arrangement of actin may for instance be disrupted.
  • the “alteration of the supramolecular arrangement of actin” may for instance correspond to a collapse of actin filaments and/or of actin stress fibers. It may also correspond to a stabilization of F-actin.
  • Altering of the supramolecular arrangement of actin may also mean enhancing the assembly of monomeric globular actin (G-actin) subunits into polymeric fibrillar actin (F-actin) filaments, or shifting the dynamic equilibrium between the G-actin and F-actin states towards F-actin.
  • G-actin monomeric globular actin
  • F-actin polymeric fibrillar actin
  • “Altering of the supramolecular arrangement of actin” may alternatively mean interfering with the formation of the actin three-dimensional architecture, for instance by altering the interaction with actin-binding proteins (ABPs).
  • an individual in need thereof refers to an individual suffering from any disease or from any affection, the mechanism of which progression and/or onset implies the integrity of the supramolecular arrangement of actin.
  • the individual to be treated in the frame of the invention is preferably a mammal, human or non human. It may also be a rodent, a feline, a canine, a bovine, an equine or an ovine. Preferably, the individual to be treated is a human being.
  • the integrity of the supramolecular arrangement of actin is required for most of the actin functions such as controlling cell shape, cytokinesis, cell motility, cell adhesion and gene expression.
  • an individual in need thereof may refer to an individual suffering from a viral or bacterial infection.
  • the integrity of the supramolecular arrangement of actin is also required for the osteoclasts function.
  • the osteoclast is the specialized cell that is responsible for bone resorption. It is a highly polarized cell that must adhere to the bone surface, where it undergoes alternative cycles of migration and resorption. Actin reorganization is critical for both processes. Osteoclast motility is mediated by podosomes, which are highly dynamic F-actin structures. Resorbing osteoclasts form a related actin complex, the sealing zone, which provides the boundary for the resorptive microenvironment. Similar to podosomes, the sealing zone is highly dependent on actin dynamics to allow efficient resorption.
  • an individual in need thereof may refer to an individual suffering from osteoporosis.
  • osteoporosis refers to any bone disease that decreases bone density and/or deteriorates bone structure.
  • compound preferably alters the osteoclastic actin ring, the podosome, and/or the sealing zone.
  • Alteration of the osteoclastic actin ring, the podosome, and/or the sealing zone may be shown by various assays well known by the skilled in the art.
  • alteration of the osteoclastic actin ring, the podosome, and/or the sealing zone may be visualized by fluorescence microscopy using e.g. phalloidin conjugated to FITC (Sigma) or Lifeact-mGFP to label F-actin.
  • Such an alteration may be visualized by electron microscopy using immuno-gold-labeling of actin.
  • Actin functions such as controlling cytokinesis, cell motility, and cell adhesion are also essential for the development of cancer and for the apparition of metastasis. Therefore, altering the supramolecular arrangement of actin may also prevent the apparition and/or development of cancer tumors.
  • an individual in need thereof refers to an individual suffering from cancer and/or metastasis.
  • the term "cancer” refers to any type of malignant (i.e. non benign) tumor.
  • the malignant tumor may correspond to a primary tumor or to a secondary tumor (i.e. a metastasis).
  • the tumor may correspond to a solid malignant tumor, which includes e.g. carcinomas, adenocarcinomas, sarcomas, melanomas, mesotheliomas, blastomas, or to a blood cancer such as leukaemias, lymphomas and myelomas.
  • the cancer is not a bone tumor or cancer.
  • the cancer is a hyperproliferative and/or an invasive cancer.
  • a hyperproliferative cancer refers to a fast growing cancer comprising cells that have escaped apoptosis and show a high metabolic rate
  • an invasive cancer refers to a cancer that comprises one or more secondary tumors or metastases.
  • the inventors have surprisingly found that treatment with the C-glucosidic ellagitannin vescalagin affects cellular morphology. Upon vescalagin treatment, cells have a more retracted appearance and exhibit irregular wound edges and retraction fibers, indicative of cell contraction (see Example 2).
  • the C- glucosidic ellagitannin compound for use for altering the supramolecular arrangement of actin in an individual suffering from cancer inhibits cell adhesion and/or cell migration.
  • the inhibition of cell adhesion and/or cell migration may be tested by various assays well known by the skilled in the art. For instance, the inhibition of cell adhesion and/or cell migration may be tested by scoring cell adhesion over time, by tracking cells under a videomicroscope, by using the wound assay or by using a transwell assay.
  • Vescalagin treatment also affects cellular viability: the inventors have shown that vescalagin treatment at 100 ⁇ for 24 hours leads to irreversible commitment to cell death (see Example 2). Therefore, in a preferred embodiment, the C-glucosidic ellagitannin compound for use for altering the supramolecular arrangement of actin in an individual suffering from cancer induces cell death.
  • the expression "cell death” may refer to any kind of cellular death such as e.g. apoptosis or necrosis. Apoptosis and necrosis may for instance be quantified by several assays well-known by the skilled in the art such as e.g. annexin V staining.
  • Apoptosis may also be specifically quantified by TUNEL assay, apoptosis-DNA ladder assay, appearance of pro-apototic markers or disappearance of anti-apoptotic markers and necrosis may for instance be discriminated from apoptosis by Propidium Iodide (PI) staining, a high PI staining being representative of necrosis.
  • PI Propidium Iodide
  • compositions according to the invention are provided.
  • the invention further pertains to a pharmaceutical composition
  • a pharmaceutical composition comprising a C- glucosidic ellagitannin and one or more physiologically acceptable carriers.
  • compositions comprising a C-glucosidic ellagitannin of the invention include all compositions wherein the C-glucosidic ellagitannin is contained in an amount effective to achieve the intended purpose.
  • the pharmaceutical compositions may contain suitable physiologically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically.
  • physiologically acceptable carrier is meant to encompass any carrier, which does not interfere with the effectiveness of the biological activity of the active ingredient and that is not toxic to the host to which is administered. Suitable physiologically acceptable carriers are well known in the art and are described for example in Remington's Pharmaceutical Sciences (Mack Publishing Company, Easton, USA, 1985), which is a standard reference text in this field.
  • the above active ingredients may be formulated in unit dosage form for injection in vehicles such as saline, dextrose solution, serum albumin and Ringer's solution.
  • compositions of the invention can also comprise minor amounts of additives, such as stabilizers, excipients, buffers and preservatives.
  • the composition of the invention may further comprise a second active principle.
  • an effective amount is meant an amount sufficient to achieve a concentration of C- glucosidic ellagitannin which is capable of preventing, treating or slowing down the disease to be treated. Such concentrations can be routinely determined by those of skilled in the art.
  • the amount of the C-glucosidic ellagitannin compound actually administered will typically be determined by a physician or a veterinarian, in the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound administered, the age, weight, and response of the subject, the severity of the subject's symptoms, and the like. It will also be appreciated by those of skilled in the art that the dosage may be dependent on the stability of the administered C- glucosidic ellagitannin.
  • Dosages to be administered depend on individual needs, on the desired effect and the chosen route of administration. It is understood that the dosage administered will be dependent upon the age, sex, health, and weight of the recipient, concurrent treatment, if any, frequency of treatment, and the nature of the effect desired.
  • the total dose required for each treatment may be administered by multiple doses or in a single dose.
  • the C-glucosidic ellagitannin of the present invention may be administered by any means that achieve the intended purpose.
  • administration may be achieved by a number of different routes including, but not limited to subcutaneous, intravenous, intradermal, intramuscular, intraperitoneal, intracerebral, intrathecal, intranasal, oral, rectal, transdermal, buccal, topical, local, inhalant or subcutaneous use. Parenteral and topical routes are particularly preferred.
  • the compounds may be formulated as liquid (e.g., solutions, suspensions), solid (e.g., pills, tablets, suppositories) or semisolid (e.g., creams, gels) forms.
  • liquid e.g., solutions, suspensions
  • solid e.g., pills, tablets, suppositories
  • semisolid e.g., creams, gels
  • the C-glucosidic ellagitannin composing the pharmaceutical composition is selected from the group consisting of vescalagin, vescalin, castalagin, castalin, grandinin, roburin A, roburin B, roburin C, roburin D, roburin E ; still preferably from the group consisting of vescalagin, vescalin, castalagin, and castalin.
  • the compound or the pharmaceutical composition of the invention may not be directly delivered on the site to be treated. Therefore, in a preferred embodiment, the compound or the pharmaceutical composition of the invention is in a mixture with a molecular delivery system.
  • the expression “molecular delivery system” refers to any kind of system that increases the concentration of the compound or the pharmaceutical composition of the invention at the site to be treated.
  • the site to be treated may be one or more tumors or one or more bones.
  • Mixing the compound or the pharmaceutical composition of the invention with a molecular delivery system may assure delivery to and maintenance at the site to be treated, leading to a better concentration of the compound at the site to be treated, and thus increasing the compound efficiency.
  • Mixing the compound or the pharmaceutical composition of the invention with a molecular delivery system may also increase the compound solubility, protect the compound against degradation and/or reduce potential side effects or the compound or the pharmaceutical composition of the invention. Therefore, when the compound or the pharmaceutical composition of the invention is in a mixture with a molecular delivery system, said molecular delivery system preferably increases the compound solubility, maintains the compound on the site to be treated, protects the compound against degradation and/or increases the compound activity.
  • said molecular delivery system may also allow controlling the time when the compound is delivered or not.
  • the compound or the pharmaceutical composition of the invention may be delivered continuously during a certain period of time and then the delivery may be suspended for a certain period of time before being resumed.
  • the compound or the pharmaceutical composition of the invention may be intermittently delivered.
  • the molecular delivery system according to the invention allows spatio-temporal controlled delivery of the compound or of the pharmaceutical composition.
  • the molecular delivery system of the invention is BioChaperoneTM, a molecular delivery system commercialized by Adocia. In vitro use of C-glucosidic ellagitannins or derivates thereof
  • the inventors have surprisingly found that the C-glucosidic ellagitannins are capable of crossing the plasma membrane, and that they specifically bind polymeric F-actin, thus interfering with its function, but not with micro-tubules. Furthermore, they have found that vescalagin does not induce depolymerization of F-actin, but rather promotes the polymerization of F-actin, thus displacing the G-actin/F-actin equilibrium in favour of F- actin.
  • vescalagin may be used as a tool for the study of the cytoskeleton.
  • it may be used for investigating cytoskeleton structure and function, and the implication of actin in various biological processes, such e.g. cell motility, ruffling, cell division, contraction, cell morphology, cell stiffness and protein secretion.
  • another aspect of the invention is a C-glucosidic ellagitannin compound or a metabolite thereof, optionally detectably labeled, for in vitro use as a tool for studying cellular mechanisms involving actin. Also provided is in vitro use of a C-glucosidic ellagitannin compound or a metabolite thereof, optionally detectably labeled, as a tool for studying cellular mechanisms involving actin.
  • cellular mechanisms involving actin may correspond to any cellular mechanism that may be impaired by disruption of the actin network integrity.
  • the “cellular mechanisms involving actin” include for instance the control or regulation of cell shape, cytokinesis, cell motility, cell adhesion, gene expression and protein secretion.
  • the C-glucosidic ellagitannin compound or the metabolite thereof may also be used in vitro for inhibiting an interaction between F-actin and a compound liable to bind F-actin.
  • a compound liable to bind F-actin may be any compound that is capable of interacting with F-actin.
  • the compound liable to bind F-actin is a polypeptide.
  • inhibiting an interaction is meant preventing the binding of a molecule to another one. The inhibition of an interaction may be measured by various methods well-known by one skilled in the art. For instance, it may be measured by western blot assays, ELISA, co- immunoprecipitation (co-ip) assays, pull-down assays, crosslinking assays, label transfer approaches (FRET or HTRF assays) or yeast two-hybrid assays.
  • the skilled in the art can easily determine if a compound inhibits an interaction between F-actin and a compound liable to bind to F-actin by carrying out a competitive binding assay.
  • the present application discloses a method for synthesizing derivates of C-glucosidic ellagitannins, such as e.g. a vescalagin-FITC conjugate or a biotinylated vescalagin conjugate (see Example 4 and Figure 3). They inventors have subsequently shown that these conjugate were able to bind actin and that the vescalagin-FITC conjugate highlighted the actin cytoskeleton, and in particular filamentous actin.
  • the C-glucosidic ellagitannin compound or a metabolite thereof for in vitro use for detecting F-actin in a cell may optionally be detectably labeled.
  • the "detectably labeled compound” may for instance be conjugated to a fluorescent moiety such as e.g. fluorescein isothiocyanate (FITC), phycoerythrin (PE), allophycocyanin (APC), cyanin (Cy), Alexa Fluor (AF).
  • FITC fluorescein isothiocyanate
  • PE phycoerythrin
  • APC allophycocyanin
  • Cy cyanin
  • Alexa Fluor (AF) Alexa Fluor
  • the "detectably labeled compound” may also be a thiol derivative, a biotin conjugate or a radiolabeled variant or conjugate.
  • Vescalagin binding actin filaments it may be a useful tool for investigating the distribution of F-actin in cells by labeling vescalagin with detectable label such as fluorescent moieties and using them to stain actin filaments for light microscopy.
  • Fluorescent derivatives of vescalagin may be very useful in localizing actin filaments in living or fixed cells as well as for visualizing individual actin filaments in vitro.
  • a high- resolution technique may developed to detect F-actin at the light and electron microscopic levels by using vescalagin conjugated to the fluorophore eosin which acts as a fluorescent tag.
  • fluorescent molecules can be utilized to drive the oxidation of diaminobenzidine (DAB) to create a reaction product that can be rendered electron dense and detectable by electron microscopy.
  • DAB diaminobenzidine
  • the amount of fluorescence visualized can be used as a quantitative measure of the amount of filamentous actin there is in cells if saturating quantities of fluorescent vescalagin are used. Consequently, immunofluorescence microscopy along with microinjection of vescalagin can be used to evaluate the direct and indirect functions of cytoplasmic actin in its different stages of polymer formation. Therefore, fluorescent vescalagin may be used as an important tool in the study of actin networks at high resolution.
  • another aspect of the invention is a detectably labeled C-glucosidic ellagitannin compound or a metabolite thereof for in vitro use for detecting F-actin in a cell. Also provided is in vitro use of a detectably labeled C-glucosidic ellagitannin compound or a metabolite thereof for detecting F-actin in a cell.
  • FIG. 1 Structures of four oak-derived C-glucosidic ellagitannins.
  • Figure 2. Live imaging of BAEc expressing actin-GFP were subjected to FRAP in the absence or presence of vescalagin (100 ⁇ ).
  • a boxed region (200 x100 pixel square) was photobleached [before (t 0"), immediately after (t 38") and after photobleaching (t 98")]; and normalized fluorescence intensity in the boxed region is shown for the entire duration of the FRAP experiment. Fluorescence recovery (starts at the red dot) recorded over time reveals the rates of actin turnover within this area.
  • B. Quantitation of the results showing the immobile fraction as calculated from the difference between pre- and post- photobleaching intensities (n 6).
  • FIG. 4 Actin polymerization at its steady state in both permissive and non- permissive conditions was continued (30 min) in the presence of either alexa633- phalloidin or vescalagin-FITC. After high-speed centrifugation, F-actin stained with alexa633-phalloidin (blue) or with vescalagin-FITC (orange), but no F-actin was detected with FITC alone; b) when the fractions obtained under similar conditions were examined for actin content by SDS-PAGE, followed by Coomassie blue staining, cytochalasin D treatment yielded G-actin and F-actin in quantities similar to those obtained in the control (CT), whereas vescalagin- or phalloidin- treated samples showed a marked depletion of G-actin. B. This effect is dose-dependent. C. This effect is not altered by the presence of the FITC-bearing unit in vescalagin-FITC.
  • BAEc were incubated for 24 hours with TGF- ⁇ (which stimulates formation of podosomes), and different concentrations of vescalagin or vescalin were added either from the beginning of the TGF- ⁇ stimulation, or during the last hour of stimulation. Cells were then fixed and treated for immunofluorescence analysis. Podosomes were visualized by double staining using phalloidin (a F-actin marker) and cortactin (a podosome marker). The percentage of total cells showing podosomes was then quantified and compared to the control (cells incubated with TGF- ⁇ only).
  • A. BAEc were stimulated for 24 hours with TGF- ⁇ , and vescalagin or vescalin was added during the last hour of stimulation.
  • Vescalagin belongs to a particular group of ellagitannins, essentially occurring in plant species of only three subclasses of the Cronquist angiosperm classification (i.e., Hamamelidae, Rosidae and Dilleniidae), and which comprises a very unique series of highly hydrosoluble C-glucosidic variants in that the usual glucopyranose core is replaced by a rarely encountered-in-nature open-chain glucose resulting from the establishment of their C-aryl glucosidic bond.
  • Cronquist angiosperm classification i.e., Hamamelidae, Rosidae and Dilleniidae
  • C-glucosidic ellagitannins Another structural feature of several of these C-glucosidic ellagitannins, including vescalagin, is the presence of a nonahydroxyterphenoyl (NHTP) unit triply connected at positions 2, 3 and 5 of their glucose core ( Figure 1 ).
  • NHTP nonahydroxyterphenoyl
  • Figure 1 The inventors' initial interest in studying these C-glucosidic ellagitannins stems from the premise that the highly pre-organized medium-sized ring-containing multiple-phenol array featured by such natural products should be structurally well-suited to interfere with the construction of protein-made cellular architectures, on top of the list of which are actin filaments and microtubules.
  • the selected compounds were the two most abundant C-glucosidic ellagitannins found in the heartwood of oak species, vescalagin and its C-1 epimer castalagin, and their corresponding two minor congeners, vescalin and castalin, both lacking the hexahydroxydiphenoyl (HHDP) unit at positions 4 and 6 of the glucose core ( Figure 1 ).
  • the inventors used bovine aortic endothelial cells (BAEc), a well-characterized type of primary cells. Any of the four ellagitannins used at 50 ⁇ rapidly provoked the disappearance of the internal stress fiber network observed in control cells.
  • cytochalasin D known to inhibit F-actin polymerization
  • the perturbed actin configuration elicited by the four C-glucosidic ellagitannins appeared distinct from those induced by cytochalasin D.
  • Example 2 Vescalagin induces rapid and sustained effects on cellular morphology
  • vescalagin-induced F-actin disrupting effect seen in BAEc was also observed in fibroblast cells (baby hamster kidney cells, BHK), which also express ⁇ -actin as the main actin isoform, as well as in smooth muscle cells (A7r5), which in contrast predominantly express a-actin.
  • Vescalagin induced similar collapse of F-actin bundles and cell contraction, but with varying potencies.
  • the impact of vescalagin on the actin cytoskeleton appeared neither cell- nor actin isoform-specific, suggesting that vescalagin can affect all types of mammalian cells.
  • Vescalagin-induced dissolution of stress fibers affected cellular morphology and, eventually, viability.
  • observations made at the light microscopic level showed that cells changed their morphology from a well spread to a more retracted appearance.
  • the cells exhibited irregular wound edges and retraction fibers, indicative of cell contraction upon vescalagin treatment. Mitosis was still observed when using vescalagin at 50 ⁇ , but became impaired at 100 ⁇ .
  • vescalagin increases the immobile fraction of actin trapped into F-actin bundles and, therefore, also affects actin dynamics within these thick F-actin bundles. From these experiments, the inventors conclude that vescalagin affects both thin and thick actin fibers, F-actin bundles made of packed actin filaments being less vulnerable to the action of vescalagin than the single filament dendritic meshwork at the cell periphery.
  • Example 4 A vescalagin-FITC conjugate directly binds F-actin
  • the inventors set up an in vitro assay based on actin polymerization from a solution of Ca 2+ -actin-ATP monomers. Spontaneous polymerization occurs when Ca 2+ is replaced by Mg 2+ provided by the F-actin buffer.
  • the inventors performed high-speed centrifugation of the samples to separate the neo-formed polymers from the monomers to discriminate binding of vescalagin to either F-actin, G-actin or both.
  • the inventors used again the vescalagin-bearing fluoprobe and a fluorescent Alexa633-phalloidin to stain F-actin.
  • Actin polymerization was carried out until it reached its steady state in both permissive and non-permissive conditions ⁇ i.e., in the presence or absence of Mg 2+ -containing F-buffer). The experiment was then continued for 30 min in the presence of either vescalagin-FITC conjugate or Alexa633-phalloidin, and the formation of actin polymers was then assessed by high-speed centrifugation. The results showed colored pellets consisting in insoluble actin stained with the fluorescent compound and indicated the expected binding of phalloidin onto filamentous actin (F- actin), as well as that of the vescalagin-FITC conjugate onto that insoluble actin material. Thus, the vescalagin-FITC conjugate directly binds F-actin.
  • F- actin filamentous actin
  • Example 5 Vescalagin binds to F-actin and promotes the actin filament state
  • a surface plasmon resonance (SPR)-based analysis confirmed that vescalagin does not bind G-actin.
  • a biotinylated vescalagin conjugate was synthesized by first reacting vescalagin with octane-1 ,8-dithiol to furnish the sulfhydryl thioether 1 -deoxyvescalagin derivative ( Figure 5). This thiol was then coupled to the biotinylated maleimide linker. This coupling reaction was performed in deuterated DMSO to enable its monitoring by 1 H NMR spectroscopy.
  • Example 6 Vescalagin aggregates actin filaments into randomly organized clusters
  • the actin filament aggregation effect of vescalagin did not prevent actin polymerization and furthermore decreases the pool of G-actin.
  • the spontaneous induction of disorganized aggregates of F-actin by vescalagin would be expected to circumvent regulated actin filament elongation at filament ends, leading to a cellular environment in which there is insufficient polymerization-competent G-actin to maintain normal stress fiber turnover. Alterations of cellular G-actin levels is known to regulate the synthesis of actin and of other actin regulatory proteins.
  • vescalagin does not bind at the same site(s) as phalloidin.
  • Example 8 Compared dose responses of endothelial cells following vescalagin or vescalin treatment
  • bovine aortic endothelial cells (BAEc) were used. These cells form podosomes following a few hours of incubation with TGF- ⁇ . Cells were fixed after 24 hours of stimulation. Two protocols were tested. In the first protocol, cells were stimulated for 24 hours with TGF- ⁇ , and vescalagin or vescalin was added during the last hour of stimulation ( Figure 7A). In the second protocol, cells were stimulated for 24 hours with TGF- ⁇ , and vescalagin or vescalin was added from the beginning of the TGF- ⁇ stimulation ( Figure 7B). In both cases, cells were then fixed and treated for immunofluorescence analysis.
  • BAEc bovine aortic endothelial cells
  • Podosomes were visualized by double staining using phalloidin (a F-actin marker) and cortactin (a podosome marker). The percentage of total cells showing podosomes was then quantified and compared to the control (cells incubated with TGF- ⁇ only) ( Figures 7A and 7B).
  • vescalagin showed a stronger inhibitor activity than vescalin.
  • the less pronounced effects observed when vescalagin or vescalin was added from the beginning of the TGF- ⁇ stimulation may be explained by rapid turn-over of these compounds.

Landscapes

  • Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Epidemiology (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • General Chemical & Material Sciences (AREA)
  • Rheumatology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne un composé d'ellagitanin C-glucosidique ou un métabolite de celui-ci destiné à être utilisé pour modifier l'arrangement supramoléculaire de l'actine chez un individu souffrant d'ostéoporose, d'un cancer, d'une infection bactérienne, ou d'une infection virale. La présente invention concerne en outre des compositions pharmaceutiques comprenant un composé d'ellagitanin C-glucosidique et/ou des métabolites de celui-ci et un ou plusieurs vecteurs physiologiquement acceptables. La présente invention concerne finalement un composé d'ellagitanin C-glucosidique ou un métabolite de celui-ci, portant facultativement un marqueur détectable, pour une utilisation in vitro en tant qu'outil destiné à étudier les mécanismes cellulaires impliquant l'actine, ou à détecter la F-actine dans une cellule.
PCT/EP2012/053017 2011-02-22 2012-02-22 Composés d'ellagitanin c-glucosidique destinés à être utilisés pour modifier l'arrangement supramoléculaire de l'actine et pour le traitement de l'ostéoporose, du cancer, d'une infection bactérienne et une d'infection virale WO2012113835A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
US14/000,766 US20130324595A1 (en) 2011-02-22 2012-02-22 C-glucosidic ellagitannin compounds for use for altering the supramolecular arrangement of actin and for the treatment of osteoporosis, cancer, bacterial infection and viral infection
EP12704839.5A EP2678012A1 (fr) 2011-02-22 2012-02-22 Composés d'ellagitanin c-glucosidique destinés à être utilisés pour modifier l'arrangement supramoléculaire de l'actine et pour le traitement de l'ostéoporose, du cancer, d'une infection bactérienne et une d'infection virale

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP11305186 2011-02-22
EP11305186.6 2011-02-22

Publications (1)

Publication Number Publication Date
WO2012113835A1 true WO2012113835A1 (fr) 2012-08-30

Family

ID=43836660

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2012/053017 WO2012113835A1 (fr) 2011-02-22 2012-02-22 Composés d'ellagitanin c-glucosidique destinés à être utilisés pour modifier l'arrangement supramoléculaire de l'actine et pour le traitement de l'ostéoporose, du cancer, d'une infection bactérienne et une d'infection virale

Country Status (3)

Country Link
US (1) US20130324595A1 (fr)
EP (1) EP2678012A1 (fr)
WO (1) WO2012113835A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9962366B2 (en) 2012-06-27 2018-05-08 Amazentis Sa Enhancing autophagy or increasing longevity by administration of urolithins or precursors thereof
WO2020201076A3 (fr) * 2019-03-29 2020-11-12 Société des Produits Nestlé S.A. Compositions et méthodes d'augmentation de la fonction des lymphocytes t
US11969408B2 (en) 2017-03-08 2024-04-30 Amazentis Sa Method for improving mitophagy in subjects

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019163437A1 (fr) * 2018-02-21 2019-08-29 株式会社ダイセル Inhibiteur de différenciation d'ostéoclastes contenant de l'urolithine
WO2019163176A1 (fr) * 2018-02-21 2019-08-29 株式会社ダイセル Inhibiteur de la différenciation des ostéoclastes contenant de l'urolithine
CN114727980A (zh) * 2019-11-27 2022-07-08 雀巢产品有限公司 苯并香豆素ampk活化剂化合物、组合物、方法及其用途

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007127263A2 (fr) * 2006-04-26 2007-11-08 The Regents Of The University Of California Utilisations thérapeutiques d'urolithines
WO2009114810A2 (fr) * 2008-03-14 2009-09-17 The Florida International Uinversity Board Of Trustees Utilisation d'ellagitannines comme inhibiteurs de la détection du quorum bactérien
US20110039796A1 (en) * 2009-04-17 2011-02-17 Zhijun Liu Natural Composition for Anti-Angiogenesis and Anti-Obesity

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007127263A2 (fr) * 2006-04-26 2007-11-08 The Regents Of The University Of California Utilisations thérapeutiques d'urolithines
WO2009114810A2 (fr) * 2008-03-14 2009-09-17 The Florida International Uinversity Board Of Trustees Utilisation d'ellagitannines comme inhibiteurs de la détection du quorum bactérien
US20110039796A1 (en) * 2009-04-17 2011-02-17 Zhijun Liu Natural Composition for Anti-Angiogenesis and Anti-Obesity

Non-Patent Citations (15)

* Cited by examiner, † Cited by third party
Title
"Remington's Pharmaceutical Sciences", 1985, MACK PUBLISHING COMPANY
BIOCHEM BIOPHYS RES COMMUN., vol. 325, no. 4, 2004, pages 1472 - 80
BONE, vol. 39, no. 5, 2006, pages 3 - 40
CHEM PHARM BULL., vol. 43, no. 11, 1995, pages 2033 - 5
HO H H ET AL: "Anti-metastasis effects of gallic acid on gastric cancer cells involves inhibition of NF-kappaB activity and downregulation of PI3K/AKT/small GTPase signals", FOOD AND CHEMICAL TOXICOLOGY, PERGAMON, GB, vol. 48, no. 8-9, 1 August 2010 (2010-08-01), pages 2508 - 2516, XP027166468, ISSN: 0278-6915, [retrieved on 20100618] *
HOFFMANN O ET AL: "Potent inhibitory effect of naturally occurring ellagitannins on in vitro osteoclastic bone resorption", BONE, PERGAMON PRESS., OXFORD, GB, vol. 39, no. 5, 1 November 2006 (2006-11-01), pages S9 - S10, XP009147298, ISSN: 8756-3282 *
ITO H: "Oxidized ellagitannins in medicinal plants and their biological activities", NATURAL MEDICINES - SHOYAKUGAKU ZASSHI, JAPANESE SOCIETY OF PHARMACOGNOSY, TOKYO, JP, vol. 59, no. 2, 1 April 2005 (2005-04-01), pages 57 - 62, XP009147315, ISSN: 1340-3443 *
ITO HIDEYUKI ET AL: "Anti-tumor promoting activity of polyphenols from Cowania mexicana and Coleogyne ramosissima", CANCER LETTERS, NEW YORK, NY, US, vol. 143, no. 1, 23 August 1999 (1999-08-23), pages 5 - 13, XP009147210, ISSN: 0304-3835, [retrieved on 19990723], DOI: DOI:10.1016/S0304-3835(99)00160-3 *
KASHIWADA Y ET AL: "ANTITUMOR AGENTS, 129. TANNINS AND RELATED COMPOUNDS AS SELECTIVE CYTOTOXIC AGENTS", JOURNAL OF NATURAL PRODUCTS, AMERICAN CHEMICAL SOCIETY, US, vol. 55, no. 8, 1 August 1992 (1992-08-01), pages 1033 - 1043, XP009064352, ISSN: 0163-3864, DOI: DOI:10.1021/NP50086A002 *
MARZOUK MOHAMED S A ET AL: "Anticancer and antioxidant Tannins from Pimenta dioica leaves", ZEITSCHRIFT FUER NATURFORSCHUNG. C, A JOURNAL OF BIOSCIENCES, TUEBINGEN, DE, vol. 62, no. 7-8, 1 July 2007 (2007-07-01), pages 526 - 536, XP009147300, ISSN: 0939-5075 *
NIRALA S K ET AL: "530 INTERCELLULAR COMMUNICATION THROUGH TUNNELING NANOTUBES: A POSSIBLE TARGET FOR ANTI-CANCER THERAPIES", JOURNAL OF HEPATOLOGY, MUNKSGAARD INTERNATIONAL PUBLISHERS, COPENHAGEN, DK, vol. 50, 1 April 2009 (2009-04-01), pages S197, XP026496143, ISSN: 0168-8278, [retrieved on 20090401], DOI: DOI:10.1016/S0168-8278(09)60532-8 *
OKUDA ET AL: "Systematics and health effects of chemically distinct tannins in medicinal plants", PHYTOCHEMISTRY, PERGAMON PRESS, GB, vol. 66, no. 17, 1 September 2005 (2005-09-01), pages 2012 - 2031, XP005076919, ISSN: 0031-9422, DOI: DOI:10.1016/J.PHYTOCHEM.2005.04.023 *
QUIDEAU ET AL., CHEM. EUR. J., vol. 11, 2005, pages 6503 - 6513
TAGURI TOSHITSUGU ET AL: "Antimicrobial activity of 10 different plant polyphenols against bacteria causing food-borne disease.", BIOLOGICAL & PHARMACEUTICAL BULLETIN DEC 2004 LNKD- PUBMED:15577214, vol. 27, no. 12, December 2004 (2004-12-01), pages 1965 - 1969, XP002633776, ISSN: 0918-6158 *
YOSHIMURA M ET AL: "Flavonol glucuronides and C-glucosidic ellagitannins from Melaleuca squarrosa", PHYTOCHEMISTRY, PERGAMON PRESS, GB, vol. 69, no. 18, 1 December 2008 (2008-12-01), pages 3062 - 3069, XP025744310, ISSN: 0031-9422, [retrieved on 20080522], DOI: DOI:10.1016/J.PHYTOCHEM.2008.04.004 *

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9962366B2 (en) 2012-06-27 2018-05-08 Amazentis Sa Enhancing autophagy or increasing longevity by administration of urolithins or precursors thereof
AU2018200791B2 (en) * 2012-06-27 2019-10-31 Amazentis Sa Enhancing autophagy or increasing longevity by administration of urolithins or precursors thereof
AU2018202407B2 (en) * 2012-06-27 2019-11-21 Amazentis Sa Enhancing autophagy or increasing longevity by administration of urolithins or precursors thereof
US11020373B2 (en) 2012-06-27 2021-06-01 Amazentis Sa Enhancing autophagy or increasing longevity by administration of urolithins or precursors thereof
US11931336B2 (en) 2012-06-27 2024-03-19 Amazentis Sa Enhancing autophagy or increasing longevity by administration of urolithins
US11931335B2 (en) 2012-06-27 2024-03-19 Amazentis Sa Enhancing autophagy or increasing longevity by administration of urolithins or precursors thereof
US11969408B2 (en) 2017-03-08 2024-04-30 Amazentis Sa Method for improving mitophagy in subjects
WO2020201076A3 (fr) * 2019-03-29 2020-11-12 Société des Produits Nestlé S.A. Compositions et méthodes d'augmentation de la fonction des lymphocytes t
CN113613663A (zh) * 2019-03-29 2021-11-05 雀巢产品有限公司 用于增加t细胞功能的组合物和方法

Also Published As

Publication number Publication date
US20130324595A1 (en) 2013-12-05
EP2678012A1 (fr) 2014-01-01

Similar Documents

Publication Publication Date Title
US20130324595A1 (en) C-glucosidic ellagitannin compounds for use for altering the supramolecular arrangement of actin and for the treatment of osteoporosis, cancer, bacterial infection and viral infection
Alexandre et al. Anticancer drug-induced cardiac rhythm disorders: current knowledge and basic underlying mechanisms
JP6603149B2 (ja) 心血管疾患を処置するためのニューレギュリン法および組成物
DE69818987T2 (de) Zusammensetzung und verfahren zur verzögerung des transports durch biologische membranen
Broers et al. Nuclear lamins: laminopathies and their role in premature ageing
TW201841936A (zh) 胜肽化合物
AU2011272137B2 (en) Novel peptide and use thereof
Sun et al. EGR1 promotes the cartilage degeneration and hypertrophy by activating the Krüppel-like factor 5 and β-catenin signaling
KR20090090315A (ko) 메타스틴 유도체 및 그의 용도
Benoist et al. The Procognitive and Synaptogenic Effects of Angiotensin IV–Derived Peptides Are Dependent on Activation of the Hepatocyte Growth Factor/c-Met System
Wan et al. Artesunate protects against surgery-induced knee arthrofibrosis by activating Beclin-1-mediated autophagy via inhibition of mTOR signaling
US20130029901A1 (en) Methods and compounds for the targeted delivery of agents to bone for interaction therewith
CN106999540A (zh) 组合疗法
US20230226140A1 (en) Ang (1-7) derviative oligopeptides for the treatment of pain
Song et al. Stretching magnitude–dependent inactivation of AKT by ROS led to enhanced p53 mitochondrial translocation and myoblast apoptosis
Batan et al. Hydrogel cultures reveal Transient Receptor Potential Vanilloid 4 regulation of myofibroblast activation and proliferation in valvular interstitial cells
US20240059685A1 (en) Piezo1 agonists for the promotion of bone formation
US20220233578A1 (en) Compositions and methods of using same for tissue regeneration
JPWO2008139952A1 (ja) 微小管破壊剤及びそれを含有する癌細胞増殖抑制剤
US20200330550A1 (en) Method for modulating insulin-independent glucose transport using teneurin c-terminal associated peptide (tcap)
KR20110007978A (ko) 콜포신 다로페이트를 포함하는 골 질환의 예방 또는 치료용 조성물
Xu et al. Targeted transplantation of engineered mitochondrial compound promotes functional recovery after spinal cord injury by enhancing macrophage phagocytosis
US20160095845A1 (en) Composition comprising (s)-(-)-benproperine for preventing or treating cancer
Tang et al. p53 peptide prevents LITAF-induced TNF-alpha-mediated mouse lung lesions and endotoxic shock
Ozaki et al. Distribution of Fos-and Jun-related proteins and activator protein-1 composite factors in mouse brain induced by neuroleptics

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 12704839

Country of ref document: EP

Kind code of ref document: A1

REEP Request for entry into the european phase

Ref document number: 2012704839

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 2012704839

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 14000766

Country of ref document: US

NENP Non-entry into the national phase

Ref country code: DE